• Title/Summary/Keyword: treatment strategy

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The Proteasome Inhibitor MG132 Sensitizes Lung Cancer Cells to TRAIL-induced Apoptosis by Inhibiting NF-κ Activation (폐암세포주에서 NFκ 활성 억제를 통한 Proteasome 억제제 MG132의 TRAIL-유도성 Apoptosis 감작 효과)

  • Seo, Pil Won;Lee, Kye Young
    • Tuberculosis and Respiratory Diseases
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    • v.65 no.6
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    • pp.476-486
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    • 2008
  • Background: TRAIL (TNF-related apoptosis inducing ligand) is a newly identified member of the TNF gene family which appears to have tumor-selective cytotoxicity due to the distinct decoy receptor system. TRAIL has direct access to caspase machinery and induces apoptosis regardless of p53 phenotype. Therefore, TRAIL has a therapeutic potential in lung cancer which frequently harbors p53 mutation in more than 50% of cases. However, it was shown that TRAIL also could activates $NF-{\kappa}B$ in some cell lines which might inhibit TRAIL-induced apoptosis. This study was designed to investigate whether TRAIL can activate $NF-{\kappa}B$ in lung cancer cell lines relatively resistant to TRAIL-induced apoptosis and inhibition of $NF-{\kappa}B$ activation using proteasome inhibitor MG132 which blocks $I{\kappa}B{\alpha}$ degradation can sensitize lung cancer cells to TRAIL-induced apoptosis. Methods: A549 (wt p53) and NCI-H1299 (null p53) lung cancer cells were used and cell viability test was done by MTT assay. Apoptosis was confirmed with Annexin V assay followed by FACS analysis. To study $NF-{\kappa}B$-dependent transcriptional activation, a luciferase reporter gene assay was used after making A549 and NCI-H1299 cells stably transfected with IgG ${\kappa}-NF-{\kappa}B$ luciferase construct. To investigate DNA binding of $NF-{\kappa}B$ activated by TRAIL, electromobility shift assay was used and supershift assay was done using anti-p65 antibody. Western blot was done for the study of $I{\kappa}B{\alpha}$ degradation. Results: A549 and NCI-H1299 cells were relatively resistant to TRAIL-induced apoptosis showing only 20~30% cell death even at the concentration 100 ng/ml, but MG132 ($3{\mu}M$) pre-treatment 1 hour prior to TRAIL addition greatly increased cell death more than 80%. Luciferase assay showed TRAIL-induced $NF-{\kappa}B$ transcriptional activity in both cell lines. Electromobility shift assay demonstrated DNA binding complex of $NF-{\kappa}B$ activated by TRAIL and supershift with p65 antibody. $I{\kappa}B{\alpha}$ degradation was proven by western blot. MG132 completely blocked both TRAIL-induced $NF-{\kappa}B$ dependent luciferase activity and DNA binding of $NF-{\kappa}B$. Conclusion: This results suggest that inhibition of $NF-{\kappa}B$ can be a potentially useful strategy to enhance TRAIL-induced tumor cell killing in lung cancer.

The Results of Radiotherapy in Locally Advanced, Unresectable Pancreatic Cancer (절제 불가능한 국소 진행된 췌장암에서 방사선치료의 결과분석)

  • Jang, Hyun-Soo;Kang, Seung-Hee;Kim, Sang-Won;Chun, Mi-Son;Jo, Sun-Mi;Lim, Jun-Chul;Oh, Young-Taek;Kang, Seok-Yun
    • Radiation Oncology Journal
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    • v.27 no.3
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    • pp.145-152
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    • 2009
  • Purpose: We retrospectively studied the outcomes and prognostic factors of patients with locally advanced, unresectable pancreatic cancer who were treated with concurrent chemoradiotherapy (CCRT) or radiotherapy only. Materials and Methods: Fifty-one patients with locally advanced, unresectable pancreatic cancer (stage IIA~III) who recevied radiotherapy ($\geq$30 Gy) between January 1994 and August 2008 were reviewed retrospectively. The median radiation dose was 39 Gy. Chemotherapy consisted of gemcitabine, cisplatin, or 5-FU alone or in various combinations, and was administered concurrently with radiotherapy in 38 patients. Results: The follow-up period ranged from 2~40 months (median, 8 months). The median survival, and the 1-and 2-year overall survival (OS) rates were 7 months, 15.7%, and 5.9%, respectively. Based on univariate analysis, the baseline CA19-9, performance status, and chemotherapy regimen were significant prognostic factors. The median survival was 8 months for CCRT, and 6 months for radiotherapy alone. The patients treated with gemcitabine-containing regimens had longer survival (median, 10 months) than the patients treated with radiotherapy alone (p=0.027). Twenty-three patients were available to evaluate the patterns of failure. Distant metastases (DM) occured in 18 patients and regional recurrences were demonstrated in 4 patients. Local progression developed in 14 patients. We analyzed the association between the time-to-DM and the baseline CA19-9 levels for 18 evaluable patients. The median time-to-DM was 20 months for patients with normal baseline CA19-9 levels and 2 months for patients with baseline CA19-9 levels $\geq$200 U/ml. Conclusion: CCRT with gemcitabine-based regimens was effective in improving OS in patients with locally advanced, unresectable pancreatic cancer. We suggest that the baseline CA19-9 level is valuable in determining the treatment strategy for patients with locally advanced, unresectable pancreatic cancer.

Germination of Two Rice Cultivars and Several Weed Species (벼와 수종(數種) 논잡초(雜草)의 휴면성(休眠性)과 발아성(發芽性))

  • Kim, Soon-Chul;Moody, Keith
    • Korean Journal of Weed Science
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    • v.9 no.2
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    • pp.116-122
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    • 1989
  • An experiment was carried out at the International Rice Research Institute in 1987 to understand the seed dormancy and germination habit of rice and several weed species. The germinability of the weed seeds just after harvest was variable depending on the species and ranged from 0 to 72%. Two rice cultivars, IR64(lowland type) and UPLRi-5(upland type) had higher than 95% in germination ability throughout the experimental period due to the fact that the rice seeds came from the harvest of the previous season and dormancy had already been overcome. The length of the storage period needed to overcome dormancy at room temperature($25{\pm}2^{\circ}C$) was about 50 days for Echinochloa glabrescens Munro ex Hook, f., more than 60 days for E. crus-galli ssp. hispidula (Retz.) Honda and 20 days for Ludwigia octovalvis(Jacq.) Raven. Seeds of E. colona(L.) Link, Monochoria vaginalis(Burm. f.) Presl, Fimbristylis miliacea(L.) Vahl and Cyperus difformis L. appeared to have no dormancy. Among the nine species M. vaginalis had the lowest germination of less than 1% throughout the experimental period. However, its seed germinated easily when planted in soil. The low germinability of E. glabrescens, E. crus-galli ssp. hispidula and L. octovalvis just after harvest could be overcome through pretreatment of seeds either by soaking in nitric acid(0.1N) for 1 day or removal of the hull in the grass species, the nitric acid treatment being superior. The results imply that germination habit of weed species varied depending on the species through their differential dormancy period or differential germination strategy.

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Mechanism Underlying a Proteasome Inhibitor, Lactacystin-Induced Apoptosis on SCC25 Human Tongue Squamous Cell Carcinoma Cells (사람혀편평상피세포암종세포에서 proteasome 억제제인 lactacystin에 의해 유도된 세포자멸사의 기전에 대한 연구)

  • Baek, Chul-Jung;Kim, Gyoo-Cheon;Kim, In-Ryoung;Lee, Seung-Eun;Kwak, Hyun-Ho;Park, Bong-Soo;Tae, Il-Ho;Ko, Myung-Yun;Ahn, Yong-Woo
    • Journal of Oral Medicine and Pain
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    • v.34 no.3
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    • pp.261-276
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    • 2009
  • Lactacystin, a microbial natural product synthesized by Streptomyces, has been commonly used as a selective proteasome inhibitor in many studies. Proteasome inhibitors is known to be preventing the proliferation of cancer cells in vivo as well as in vitro. Furthermore, proteasome inhibitors, as single or combined with other anticancer agents, are suggested as a new class of potential anticancer agents. This study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying induction of apoptosis in SCC25 human tongue sqaumous cell carcinoma cell line treated with lactacystin. The viability of SCC25 cells, human normal keratinocytes (HaCaT cells) and human gingiva fibroblasts (HGF-1 cells), and the growth inhibition of SCC25 cells were assessed by MTT assay and clonogenic assay respectively. The hoechst staining, hemacolor staining and TUNEL staining were conducted to observe SCC25 cells undergoing apoptosis. SCC25 cells were treated with lactacystin, and Western blotting, immunocytochemistry, confocal microscopy, FAScan flow cytometry, MMP activity, and proteasome activity were performed. Lactacystin treatment of SCC25 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. Interestingly, lactacytin remarkably revealed cytotoxicity in SCC25 cells but not normal cells. And tested SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA contents, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, the up-regulation of Bax, and the activation of caspase-7, caspase-3, PARP, lamin A/C and DFF45 (ICAD). Flow cytometric analysis revealed that lactacystin resulted in G1 arrest in cell cycle progression which was associated with up-regulation in the protein expression of CDK inhibitors, $p21^{WAF1/CIP1}$ and $p27^{KIP1}$. We presented data indicating that lactacystin induces G1 cell cycle arrest and apoptois via proteasome, mitochondria and caspase pathway in SCC25 cells. Therefore our data provide the possibility that lactacystin could be as a novel therapeutic strategy for human tongue squamous cell carcinoma.

Cryopreservation of Primordial Germ Cells(PGCs) from Korean Native Chicken(Ogye) Embryos using Commercial Cryoprotectants (상업용 동결보호제를 이용한 한국재래닭(오계) 원시생식세포의 동결 보존)

  • Kim, Hyun;Kim, Dong Hun;Han, Jae Yong;Do, Yoon Jung;Kim, Jae Hwan;Kim, Young Sin;Seong, Hwan Hoo;Ko, Yeoung Gyu;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.163-169
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    • 2013
  • Cryopreservation of poultry semen has been reported, but preservation of female genetic material has not been possible because of the unique anatomical and physiological characteristics of the avian egg. Thus an alternative strategy for conservation of oviparous species of animals must be developed. Recent technological developments for producing germline chimeras by the transfer of primordial germ cells (PGCs) into recipient embryos has enabled the conservation and retrieval of chicken genetic resources in their complete form. In the present study, fertilized eggs were incubated for about 5.5 days to obtain embryos at stage 28. The whole embryo was collected from the germinal gonad using a fine glass micro pipette under a microscope. The PGCs were then purified using MACS method. Two commercially available cryoprotectants (A and B) were used to preserve the PGCs, and EG were used as a control. The average recovery rate of PGCs after thawing was 35.5% and 60.5% with the A and B treatments, respectively. There was no significant difference between B treatments and control, which showed an average recovery rate of 52.8%. However, the recovery rate obtained using A cryoprotectant (35.5%) was significantly lower than using treatment control and B. The average viability of the PGCs after thawing were 77.9% and 77.4% for cryoprotectants A and B, respectively, and the control were was 81.6%. There was no statistically significant difference between the two treatments and control. It was concluded that all of the available cryoprotectants examined in this study could be used for preservation of PGCs from embryos. Further experiments to produce germline chimera from PGCs preserved using this techniques are strongly recommended.

The Level of Diabetes Management of Agriculture, Forestry, and Fishery Workers (농림어업인의 당뇨병 관리 수준)

  • Oh, Gyung-Jae;Lee, Young-Hoon
    • Journal of agricultural medicine and community health
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    • v.42 no.3
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    • pp.119-131
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    • 2017
  • Objectives: The purpose of this study was to compare the diabetic management indicators between agriculture, forestry, and fishery workers (AFF) and other occupational adults (non-AFF) in community-dwelling diabetes. Methods: The study population consisted of 22,127 diabetic population ${\geq}19years$ who participated in the 2015 Community Health Survey. Chi-square test and logistic regression analysis was used to compare the diabetic management indicators between AFF and non-AFF. Socioeconomic characteristics such as age, gender, education level, monthly household income, National Basic Livelihood Security status, and marital status was sequentially adjusted. Results: Among total diabetic population, 3,712 people (16.8%) was AFF and 18,415 people (83.2%) was non-AFF. The fully-adjusted odds ratio [OR] (95% confidence interval [CI]) of current non-medical treatment (0.72, 0.66-0.79), measurement of hemoglobin A1c (0.61, 0.55-0.67), screening for diabetic retinopathy (0.76, 0.70-0.83), screening for diabetic nephropathy (0.75, 0.70-0.81), non-alcoholic or moderate drinking (0.70, 0.64-0.78), nutrition label reading (0.83, 0.71-0.98), low salt preference (0.85, 0.78-0.93), dental examination (0.60, 0.54-0.66), scaling experience (0.84, 0.77-0.93), regular toothbrushing (0.66, 0.58-0.76), and diabetes management education (0.84, 0.77-0.92) was significantly lower in AFF compared to non-AFF. In contrast, the fully-adjusted OR (95% CI) of AFF's low stress level (1.39, 1.26-1.52) and adequate sleep duration (1.22, 1.13-1.32) was significantly higher than non-AFF, which are better indicators of diabetic management in AFF. Conclusions: Overall, the level of diabetes management of AFF was not as good as that of non-AFF. In order to improve the level of diabetes management of AFF, a delicate diabetes intervention strategy considering the occupational characteristics of AFF will be needed.

Suppression of Helicobacter pylori-induced Angiogenesis by a Gastric Proton Pump Inhibitor (Proton Pump Inhibitor에 의한 Helicobacter pylori의 혈관형성 억제효과)

  • Jin, Sung-Ho;Lee, Hwa-Young;Kim, Dong-Kyu;Cho, Yong-Kwan;Hahm, Ki-Baik;Han, Sang-Uk
    • Journal of Gastric Cancer
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    • v.5 no.3 s.19
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    • pp.191-199
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    • 2005
  • Background: Though infections of Helicobacter pylori (H. pylori) are closely associated with activation of host angiogenesis, the underlying mechanisms, as well as the strategy for its prevention, have not been identified. Here, we investigated a causal role of H. pylori infection in angiogenesis of gastric mucosa and a potent inhibitory effect of a gastric proton pump inhibitor (PPI) on the gastropathy. Materials and Methods: A comparative analysis of CD 34 expression in tissues obtained from 20 H. pylori-associated gastritis and 18 H. pylori-negative gastritis patients was performed. Expression of $HIF-1{\alpha}$ and VEGF were tested by using RT-PCR. To evaluate the direct effect of H. pylori infection on differentiation of endothelial HUVEC cells, we carried out an in vitro angiogenesis assay. Results: H. pyfori-associated gastritis tissues showed significantly higher density of $CD34^+$ blood vessels than did H. pylori-negative gastritis tissues, and the levels were well correlated with expressions of $HIF-1{\alpha}$. Conditioned media from H. pylori-infected gastric mucosal cells stimulated a tubular formation of HUVEC cells. We also found a significant inhibitory effect of PPI, an agent frequently used for H. pylori eradication, on H. pylori-induced angiogenesis. This drug effectively inhibited the phosphorylation of MAP kinase ERK1/2, which is a principal signal for H. pylori-induced angiogenesis. Conclusion: The fact that PPls can down-regulate H. pylori-induced angiogenesis suggest that anti-angiogenic treatment using PPI may be a preventive approach for H. pylori-associated carcinogenesis.

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TIMP-2 Gene Transfer Via Adenovirus Inhibits the Invasion of Lung Cancer Cell (TIMP-2 유전자 재조합 아데노바이러스의 폐암세포 침윤 억제 효과)

  • Oh, Yeon-Mok;Lee, Jae-Ho;Yoo, Chul-Gyu;Chung, Hee-Soon;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.2
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    • pp.189-197
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    • 2000
  • Background : Tissue inhibitor of metalloproteinase is a natural inhibitor that counteracts pro teolytic enzymes essential to the invasion of cancer cell. Whether or not TIMP-2 gene transfer via adenovirus could inhibit the invasion of lung cancer cell iη vitro was evaluated for the future purpose of gene therapy against lung cancer. Methods : Recombinant adenovirus-TIMP-2(Ad-TIMP-2) was generated by homologous recombination after pACCMV-TIMP-2 and pJM17 were cotransfected into 293 cell by standard calcium phosphate coprecipitate method. Calu-6, one of the most invasive lung cancer cells, was transduced with Ad-TIMP-2 or Ad-$\beta$gal. Anchorage-independent growth and invasiveness were assessed by soft agar clonogenicity assay and invasion assay using two-chamber, well divided by matrigel. Results : Ad-TIMP-2 transduced calu-6 cells produced biologically active TIMP-2 more than 50 times more than parental calu-6. TIMP-2 gene transfer did not suppress the in vitro tumorigenicity. However, two chamber well assay revealed that Ad-TIMP-2 transduction reduced the invasiveness of calu-6 efficiently (12% compared with parental cell) even at low 10moi. Conclusion : Even though TIMP-2 gene transfer did not inhibit in vitro tumorigenicity, it did inhibit invasion of lung cancer cell in vitro. The inhibition of invasion by Ad-TIMP-2 may be a useful strategy for the treatment of lung cancer.

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Post-operative Radiation Therapy for Esophageal Cancer; Analysis of Failure Pattern (식도암의 수술 후 방사선 치료: 실패 양상 분석)

  • Kim Mi Sook;Kim Jae Young;Yoo Seoung Yul;Zo Chul Goo;Yoo Hyung Jun;Zo Jae Ill;Baek Hee Jong;Park Jong Ho;Choi Soo Yong
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.447-454
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    • 1998
  • Purpose : This study evaluated the survival, local control, prognostic factor, and failure pattern of patients with esophageal cancer treated with operation and adjuvant radiation therapy to use as fundermental data of postoperative radiation therapy. Materials and Methods : A retrospective analysis was undertaken of 82 patients who had locally advanced esophageal cancer treated with operation and adjuvant radiation therapy from January 1988 to December 1995. According to AJCC staging, stage IIA were in 26 patients, stage IIB in 4 patients, and stage III in 52 patients. Squamous cell carcinoma were in 77 patients, adenosquamous carcinoma in 3 patients, and adenocarcinoma in 2 patients. The patients received radiation therapy ranging from 41.0 Gy to 64.8 Gy. Five patients received neoadjuvant chemotherapy. Results : Two-year survival and local control rates for all patients were 36.8$\%$ and 30.4$\%$ respectively. And they were 9.3$\%$ and 26.3$\%$ respectively at 5 years. According to stages, 2-year survival rates were 50.2$\%$ in IIA, 0$\%$ in IIB and 23.3$\%$ in III (p=0.004). Two-year local control rates were 49.2 $\%$ in IIA, 66.6$\%$ in IIB and 24.7$\%$ in III (p=0.01). Sixty patients developed recurrence, which were 3 tumor margin, 23 lymph node recurrence, 4 tumor margin and lymph node, 1 tumor margin and distant metastasis, 9 lymph node and distant metastasis, 17 distant metastasis and 3 unknown metastatic site. Prognostic factors affecting survival were smoking (p=0.02), T-staging (p=0.0092), N-staging (p=0.0045). Prognostic factors affecting local control were T-staging (p=0.019), N-staging (p=0.047). Conclusion : In spite of post-operative radiation therapy, predominant failure pattern was local failure. Especially regional lymph node failure was major cause of local failure. So strategy of aggresive adjuvant radiation therapy to regional lymph node area in post operative treatment should be proposed.

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Establishment of Discriminating Concentration based Assessment for Insecticide Resistance Monitoring of Palm thrips (오이총채벌레의 약제 저항성 진단을 위한 판별농도 기반 생물검정법 확립)

  • Jeon, Sung-Wook;Park, Bueyong;Park, Se-Keun;Lee, Sang-Ku;Ryu, Hyun-Ju;Lee, Sang-Bum;Jeong, In-Hong
    • Korean Journal of Environmental Biology
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    • v.35 no.4
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    • pp.557-565
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    • 2017
  • For our survey of insecticidal resistance of Palm thrips (Thrips palmi Karny), we established the discriminating time (DT) and concentration (DC) of nine insecticides, and we conducted a bioassay about seven local populations via leaf-dipping methods. The discriminating times of the recommended concentration (RC) were 24 h at emamectin benzoate EC and spinetoram SC, 48 h at chlorfenapyr EC, 72 h at spinosad SC, cyantraniliprole EC, acetamiprid WP, dinotefuran WG, imidacloprid WP and thiacloprid SC after treatment. The DC estimated the concentration which showed the difference within the mortalities of these local populations. The DCs were emamectin benzoate EC $0.013mg\;L^{-1}$ (RC, $10.8mg\;L^{-1}$), spinetoram SC $0.125mg\;L^{-1}$ (RC, $25.0mg\;L^{-1}$), chlorfenapyr EC $0.25mg\;L^{-1}$ (RC, $50.0mg\;L^{-1}$), spinosad SC $0.083mg\;L^{-1}$ (RC, $50.0mg\;L^{-1}$) and cyantraniliprole EC $5.0mg\;L^{-1}$ (RC, $50.0mg\;L^{-1}$), and DCs of neonicotinoids were their RCs, that is, acetamiprid WP (RC, $40.0mg\;L^{-1}$), dinotefuran WG (RC, $20.0mg\;L^{-1}$), imidacloprid WP(RC, $50.0mg\;L^{-1}$) and thiacloprid SC (RC, $50.0mg\;L^{-1}$). From our investigation into the resistance of the local populations with DT and DC application, the neonicotinoid insecticides have shown a high resistant level for all the local populations, and the other insecticides have demonstrated low or non-resistance. In the use of neonicotinoid insecticides to control Palm thrips, one must take caution. As a result, the establishment of DT and DC in the single dose bioassay method was helpful for surveying the insecticide response dynamics and the development of an insecticide resistance management strategy.